GB/T 44342-2024 苏铁叶枯病菌检疫鉴定方法

ICS

65.020.20

CCS

B16

中华人民共和国国家标准

GB/T44342—2024

苏铁叶枯病菌检疫鉴定方法

Detectionandidentificationof

Didymellaglomerata（Corda）QianChen&L.Cai

2024-08-23发布2025-03-01实施

国家市场监督管理总局发布

国家标准化管理委员会

GB/T44342—2024

目次

前言

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范围

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2

规范性引用文件

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3

术语和定义

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4

苏铁叶枯病菌基本信息

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5

方法原理

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试剂、材料和培养基

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6.1

试剂和材料

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6.2

培养基

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仪器和用具

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7.1

仪器

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7.2

用具

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检疫鉴定方法

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8.1

病害症状

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8.2

病原菌分离培养和形态鉴定

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8.3

分子生物学鉴定

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结果判定

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10

样品和档案保存

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10.1

样品保存与处理

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10.2

菌株保存与处理

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10.3

结果记录与资料保存

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附录A（资料性）苏铁叶枯病菌的其他信息

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A.1

病害情况

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A.2

寄主范围

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A.3

分布范围

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A.4

传播方式

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A.5

病害症状

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A.6

病原菌形态特征

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附录B（资料性）苏铁叶枯病菌培养基配制方法

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B.1

马铃薯葡萄糖琼脂（PDA）培养基

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B.2

燕麦片琼胶（OA）培养基

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附录C（资料性）苏铁叶枯病菌DNA提取

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C.1

DNA提取方法（改良CTAB提取法）

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C.2

DNA纯度与浓度的测定

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Ⅰ

GB/T44342—2024

附录D（规范性）苏铁叶枯病菌实时荧光PCR检测方法

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D.1

引物和探针序列

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D.2

实时荧光PCR反应体系

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D.3

结果判定

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附录E（规范性）苏铁叶枯病菌β微管蛋白tub2片段PCR扩增方法

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E.1

引物序列

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E.2

PCR反应体系及参数

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参考文献

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GB/T44342—2024

前言

本文件按照GB/T1.1—2020《标准化工作导则第1部分：标准化文件的结构和起草规则》的规

定起草。

请注意本文件的某些内容可能涉及专利。本文件的发布机构不承担识别专利的责任。

本文件由全国植物检疫标准化技术委员会（SAC/TC271）提出并归口。

本文件起草单位：中国检验检疫科学研究院、中国科学院微生物研究所、深圳仙湖植物园、宁波海

关技术中心、福建省热带作物科学研究所、西藏自治区高原生物研究所。

本文件主要起草人：刘芳、黄英、程颖慧、蔡磊、徐晗、段维军、蔡江桥、罗金水、王聪、严进、

普布多吉。

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GB/T44342—2024

苏铁叶枯病菌检疫鉴定方法

1范围

本文件描述了苏铁叶枯病菌（Didymellaglomerata）的分离培养、形态学鉴定、分子生物学鉴定

方法。

本文件适用于植物及其产品中苏铁叶枯病菌的筛查鉴定。

2规范性引用文件

本文件没有规范性引用文件。

3术语和定义

本文件没有需要界定的术语和定义。

4苏铁叶枯病菌基本信息

学名：Didymellaglomerata（Corda）QianChen&L.Cai

异名：ConiothyriumglomeratumCorda,Icon.Fungorum（Prague）4:39.1840.

Aposphaeriaglomerata（Corda）Sacc.,Syll.Fung.（Abellini）3:175.1884.

Phomaglomerata（Corda）Wollenw.&Hochapfel,Z.Parasitenk.3（5）:592.1936.

Peyronellaeaglomerata（Corda）Goid.exTogliani,Ann.Sperim.Agrar.Ⅲ6:93.1952.

中文学名：团集亚隔孢壳

中文异名：葡萄茎枯病菌

分类地位：真菌界Fungi，子囊菌门Ascomycota，座囊菌纲Dothideomycetes，格孢腔菌目Pleospor﹘

ales，亚隔孢壳科Didymellaceae，亚隔孢壳属Didymella。

苏铁叶枯病菌与亚隔孢壳属中其他物种的形态特征相似，难以单纯依靠形态特征进行区分。近缘种

包括D.anserina、D.aurea、D.musae等。

苏铁叶枯病菌的其他信息见附录A。

5方法原理

根据病害症状、分离菌的形态特征（见附录A），以及实时荧光聚合酶链式反应（PCR）检测方法

或β微管蛋白tub2序列特征对苏铁叶枯病菌进行检测鉴定。

6试剂、材料和培养基

6.1试剂和材料

除有特殊说明外，所有试验用试剂均为分析纯或生化试剂。

分离培养所需试剂和材料：马铃薯、燕麦片、蒸馏水、葡萄糖、琼脂粉、次氯酸钠、75%乙醇、无菌水。

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